Project Summary/Abstract There exists a dire need for new and more effective drugs to treat disorders of the central nervous system (CNS), especially in mental health. The pace of approval for new chemical entities is anemic, with an average of just three compounds per year entering the market over the last 15 years. Unsurprisingly, clinical trial failure rates for CNS drug development remain greater than 90%, and many more programs fail during preclinical development. Given the immense personal, societal, and economic burden that psychiatric disorders place on the United States, better drug discovery and development paradigms must be implemented. BrainXell is addressing this issue by introducing new in vitro model systems with high relevance to the human brain that are composed of normal cultured human neurons of various types. This proposal focuses on one of these types, cortical glutamatergic neurons, which is central to many psychiatric and neurological disorders. In our Phase I project, we developed a novel and proprietary cocktail to expand cortical glutamatergic progenitors, allowing generation of 5103 glutamatergic neurons from a single induced pluripotent stem cell (iPSC). This expansion method exhibits a clear advantage over traditional approaches that use growth factors, such as fibroblast growth factor (FGF2) and/or epidermal growth factor (EGF), as glutamatergic progenitors expanded under such conditions often change their differentiation potential. We have also optimized the cell dissociation process and neuron cryopreservation medium, which yield cell viability >90% after dissociating into single cells and >70% after thawing. Together, these studies provide the technical basis to accomplish the same goal: manufacture large-scale, high-purity cortical glutamatergic neurons. This technology developed in Phase I will be fully commercialized in Phase II. We will produce multiple large lots of normal neurons to validate our protocol. Our procedures will be augmented with new equipment to ensure consistency between vials and lots. The characterization and quality control procedures for the resulting product will be optimized and expanded to provide customers with a complete data set and clear quality criteria for the neurons. Continuing our focus on product performance for our customers, we will generate ?best practice? protocols for the end user that give guidance for culturing neurons in a range of systems. Because glutamatergic neuron production will be offered as a service, we will also test our methods on outside iPSC lines derived from individuals with schizophrenia and healthy controls. Finally, we will populate our product pipeline by developing protocols to produce glutamatergic neurons specific for superficial (layer II/III) cortical layers.